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Aspergillus parasiticus and its application in the preparation of nitrite reductase, nitrite reductase gene and genetic engineering bacteria

A technology of Aspergillus parasiticus and preservation number, applied in the field of microorganisms, can solve the problems of low efficiency, secondary pollution, unsuitable for aquaculture water and food processing, etc., and achieve the effects of good degradation effect, high efficiency and low energy consumption, and fast and reliable degradation.

Inactive Publication Date: 2017-05-03
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many problems in these methods. Among them, the reverse osmosis method, electrodialysis method, and ion exchange method all concentrate nitrite in the medium or waste liquid, which is easy to cause secondary pollution, and has low efficiency and large investment. It is suitable for the treatment of industrial water; although the chemical reduction method can reduce nitrite to harmless nitrogen, the reaction needs to be carried out at a higher temperature, and the efficiency is low, so it is not suitable for the treatment of aquaculture water and food; Although the chemical oxidation method has the advantages of simple equipment and relatively low treatment costs, it is only suitable for the treatment of deep underground water with low nitrite content and less treatment capacity, and is not suitable for the treatment of aquaculture water and food

Method used

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  • Aspergillus parasiticus and its application in the preparation of nitrite reductase, nitrite reductase gene and genetic engineering bacteria
  • Aspergillus parasiticus and its application in the preparation of nitrite reductase, nitrite reductase gene and genetic engineering bacteria
  • Aspergillus parasiticus and its application in the preparation of nitrite reductase, nitrite reductase gene and genetic engineering bacteria

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, the screening of aspergillus parasitica strain JFS, culture, identification and performance test

[0032] 1. Screening of Aspergillus parasiticus strain JFS:

[0033] (1) Material

[0034] a. Samples: 20 samples of water samples, soil samples, dead wood, rabbit manure and sheep manure collected from Sichuan Chengdu, Sichuan Yibin and Chongqing Rongchang.

[0035] Sample processing and storage: The collected soil samples, rotten wood, rabbit manure and sheep manure were all dried in a cool and dry place immediately after being collected, and stored in clean kraft paper envelopes after drying. Water samples were used directly after collection (fresh water was used).

[0036] b. Medium:

[0037]

[0038]

[0039] All the above-mentioned media were prepared in 1×10 5 Under Pa, disinfect for 25 minutes.

[0040] c. Reagents and instruments:

[0041] Reagents: Potassium ferrocyanide, zinc acetate, p-aminobenzenesulfonic acid, naphthaleneethylenediami...

Embodiment 2

[0118] Embodiment 2, the application of Aspergillus parasiticus in the preparation of nitrite reductase gene

[0119] The preparation steps of nitrite reductase gene are as follows:

[0120] (a) Transfer the strain JFS from the slant of the Chase culture medium to the Chase plate or the Chase slant for 3-7 days at 30°C, or transfer to the Chase seed medium, 30°C, 150r / min After culturing for 3 to 8 days, centrifuge or filter to collect bacteria and sporophytes.

[0121] (b) Extract DNA from the collected bacteria and sporophytes: extract the genomic DNA of strain JFS.

[0122] (c) Design degenerate primers through the conserved region of the nitrite reductase amino acid sequence of various organisms in the Genbank protein database, and use the genomic DNA of the bacterial strain JFS as a template to amplify the DNA fragment of the nitrite reductase in the bacterial strain JFS, Then sequence. Existing technologies can be used for specific gene preparation, which will not be ...

Embodiment 3

[0124] Embodiment 3, the application of Aspergillus parasiticus in the preparation of nitrite reductase

[0125] The strain JFS was transferred from the slant of Chase's medium into Chase's seed medium respectively, and after culturing for 2 days at 30°C and 150r / min, it was transferred into the fermentation medium at 30°C and 150r / min with an inoculum size of 3-10%. Centrifuge or filter for 3 to 9 days, collect bacteria and sporophytes, use ultrasonic crushing or liquid nitrogen grinding or glass bead grinding or use the above three wall breaking methods to break the cell wall of strain JFS, and then extract, Purify nitrite reductase by salting out, chromatography and other methods. The specific preparation method of nitrite reductase can adopt the prior art, and will not be repeated here.

[0126] The Aspergillus parasitica of the present invention is used in the preparation of the nitrite reductase. Since the Aspergillus parasitica of the present invention is easy to culti...

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Abstract

The invention discloses an aspergillus parasiticus and its application in the preparation of nitrite reductase, nitrite reductase gene and genetic engineering bacteria containing nitrite reductase. The preservation number of aspergillus parasiticus is CCTCC M2013662. The invention provides an aspergillus parasiticus with high efficiency, low energy consumption, good effect on nitrite degradation, fast and reliable degradation. And the aspergillus parasiticus can be well applied in the preparation of nitrite reductase, nitrite reductase gene preparation and genetically engineered bacteria containing nitrite reductase.

Description

technical field [0001] The invention belongs to the technical field of microbes, relates to an aspergillus and its application, in particular to an aspergillus parasitica, and the preparation of nitrite reductase, gene preparation of nitrite reductase and the gene containing nitrite reductase of the aspergillus parasiticus Application in the preparation of engineering bacteria. Background technique [0002] Nitrite is one of the most common nitrogen-containing compounds in nature, and its sources are also very extensive. Nitrite is a harmful chemical substance. Nitrite reacts with human blood to form methemoglobin, which makes the blood lose oxygen. Function, make people hypoxic poisoning, dizziness, palpitation, vomiting, lip bruising in mild cases, confusion, convulsions, shortness of breath in severe cases, life-threatening if not rescued in time. Not only that, nitrite can also react with secondary amines in the human body to form nitrosamines, which are carcinogenic, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/06C12N15/53C12N15/70C12N15/81C12R1/66
CPCC12N9/0044C12Y107/02002C12N1/145C12R2001/66
Inventor 刘冰花杨林蒲小龙罗亚雄袁管明
Owner CHENGDU UNIV
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